Multicolour detection – Long Ex/Em wavelengths of Cal-590™ & Cal-630™ make these dyes compatible for use with green fluorescent protein (GFP) cell lines.

Features

Predominantly localised in cytosols.

Convenient, can be easily detected by standard filter sets for FITC, TRITC/Cy3 and Texas Red®

Versatile packing sizes to meet your special needs.

Cal-520™, Cal-590™ and Cal-630™ provide the most robust homogeneous
fluorescence-based assay tools for detecting intracellular calcium
mobilisation. They are fluorogenic calcium-sensitive dyes with a
significantly improved signal-to-noise ratio and intracellular retention
compared to the existing calcium indicators (such as Fluo-3 AM, Fluo-4
AM and Rhod-2 AM). Cells expressing a GPCR or calcium channel of
interest that signals through calcium can be preloaded with Cal-520™ AM, Cal-590™ AM or Cal-630™ AM which can cross the cell membrane. Once inside the cell, the lipophilic blocking groups of Cal 520™ AM, Cal-590™ AM or Cal-630™
AM are cleaved by intracellular esterases, resulting in a negatively
charged fluorescent dye that stays inside cells. Their fluorescence is
greatly enhanced upon binding to calcium. When cells are stimulated with
agonists, the receptor signals the release of intracellular calcium,
which significantly increase the fluorescence of Cal-520™, Cal-590™ or Cal-630™. The characteristics of high sensitivity and >100 times fluorescence enhancement make Cal-520™ AM, Cal-590™ AM or Cal-630™
AM ideal indicators for the measurement of cellular calcium. The high
S/N ratio and better intracellular retention make the Cal-520™, Cal-590™ or Cal-630™
calcium assay a robust tool for evaluating GPCR and calcium channel
targets as well as for screening their agonists and antagonists.

Besides their convenient excitation wavelengths and large fluorescence enhancement by calcium, Cal-520™, Cal-590™, and Cal-630™
are predominantly localised in cytosols unlike Rhod-2 and x-Rhod-1 that
are mainly localized in mitochondria. In addition, the long Ex/Em
wavelengths of Cal-590™ and Cal-630™ make these dyes perfect calcium
indicators compatible for multicolour detection with green fluorescent
protein (GFP) cell lines. In addition, Cal-520™, Cal-590™ or Cal-630™
calcium assays are optimised to be compatible with most of the existing
fluorescence instruments. Cal-520 can be well excited at 488 nm, and
used with FITC filter set. Cal-590 is optimized to be excited at 555 nm,
and used with TRITC/Cy3 filter set. Cal-590 is optimized to be excited
at 594 nm, and used with Texas Red® filter set. The spectral and calcium binding properties are summarized below (see Table 1).

Table 1. Spectral and Ca2+–Binding Properties of Cal-520™, Cal-590™

Ca2+ Indicator

Excitation (nm)

Emission (nm)

Kd (nm)

Filter Set

Cal-520™

492

514

320

FITC

Cal-590™

573

588

561

TRITC/Cy3

Cal-630™

608

626

792

Texas Red®

Product Range

Cal-520™, AM Cell-Permeable

Cal-520™ Sodium Salt

Cal-520™ Potassium Salt

Cal-520FF™, AM cell-permeable

Cal-520FF™ Potassium Salt

Cal-520-Dextran Conjugate MW 3,000

Cal-520-Dextran Conjugate MW 10,000

Cal-590™ AM Cell-permeable

Cal-590™ Sodium salt

Cal-590™ Potassium Salt

Cal-630™ AM Cell-permeable

Cal-630™ sodium salt

Cal-630™ potassium salt

Because of the importance of Ca2+ in biology, numerous
techniques/methods for analysing the mechanisms of cellular and/or
subcellular Ca2+ activity have been established. Although each method
for analysing Ca2+ activity has certain advantages of the others, each
also suffers for drawbacks. With the outstanding properties of Cal-520™, Cal-590™ & Cal-630™,
calcium detection reagents provide a new powerful tool for
intracellular calcium analysis and monitoring in a variety of biological
systems.

The interests of many researchers in Ca2+ analysis has shifted from
the cellular level to the subcellular level. It has been found that
Ca2+ is not even distributed throughout the whole cell and that
intracellular heterogeneity of Ca2+ (such as Ca2+ waves and Ca2+ sparks)
is observed in a variety of cells (e.g. oocyte, heart muscle cell,
hepatocyte, and exocrine cells). With the advent of the confocal laser
scanning microscope (CLSM) in the 1980’s and advanced miscroplate
readers in 2000’s (such as FLIPR, FDSS and NOVOStar dedicated for
intracellular Ca2+ detections), the measurement of intracellular Ca2+
has accelerated significantly. Confocal laser scanning microscopy and
more recently multiphoton microscopy allow the precise spatial and
temporal analysis of intracellular Ca2+ signalling at the subcellular
level in addition o the measurement of its concentration.

Stratech Scientific is a distributor of high quality, competitively
priced, reliable products for research laboratories throughout the UK
and Europe. Please contact us to find out which ranges we can supply in
your country.